1. Field
The present disclosure relates to a composition and a kit for detecting a nucleic acid with genetic variation, and a method of detecting a nucleic acid with genetic variation by using the same.
2. Description of the Related Art
Molecular diagnostic methods have been widely used in the fields of medicine, identification, the environment, etc. Detection of a variant of a specific nucleic acid sequence can provide information regarding polymorphism or mutations including disease-related mutations.
Numerous methods of detecting the presence of disease-related genetic variations or specific genotypes have been developed and used. Examples of the methods include direct sequencing, allele-specific PCR, Restriction Fragment Length Polymorphism (RFLP), Taqman™ probe technique, amplification refractory mutation system (ARMS)-PCR, denaturing HPLC (dHPLC), and real-time PCR, etc. Major factors considered in detecting genetic variations are sensitivity, which enables detection of mutant DNA present at a low rate in normal DNA, and specificity, which enables minimalization of the rate of false positive, i.e., the rate of normal DNA being falsely determined as mutant DNA.
However, the existing methods have not been successful in showing a high reliability level of sensitivity and specificity. For example, the direct sequencing method has the advantage of having a relatively low rate of false positives due to high specificity but it has the disadvantage of enabling detection only when at least about 20 to about 30% of mutant DNA is present in a given sample.
Thus, there is a desire for a method that is capable of detecting with high sensitivity and accuracy even when the frequency of nucleic acids with genetic variation is low.